Brown adipocyte progenitors in human skeletal muscle
Inventors
Boss, Olivier D. • Crisan, Mihaela • Giacobino, Jean-Paul
Assignees
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Publication Number
US-10301595-B2
Publication Date
2019-05-28
Expiration Date
Abstract
This invention relates to brown adipose tissue (BAT) progenitor cells and methods for isolating BAT progenitor cells from skeletal muscle. BAT progenitor cell surface markers and medium and agents for inducing cell differentiation into brown adipocytes are also provided. In some embodiments, the BAT progenitor cell expresses a first cell surface marker associated with endothelial cells, the first cell surface marker being detectable in an antibody based assay using a first antibody. In addition, the BAT progenitor cell can be substantially free of a second cell surface marker associated with endothelial cells, the second cell surface marker being substantially undetectable in said antibody based assay using a second antibody. The BAT progenitor cell can also be substantially free of additional cell surface markers.
Core Innovation
The invention provides a method for identifying a brown adipose tissue (BAT) progenitor cell from vascular cells isolated from human skeletal muscle. The method contacts the vascular cell population with a first antibody specific to a CD34 marker and a second antibody specific to a CD31 marker, and then determines, in an antibody based assay, a BAT progenitor cell that expresses the CD34 marker and is substantially free of the CD31 marker. The approach can further use antibody-based selection and exclusion to define additional cell populations that are substantially undetectable in the assay.
The invention further describes culturing BAT progenitor cells obtained from human skeletal muscle of an individual or a donor in a medium, where the BAT progenitor cells are vascular cells that express CD34 and are substantially free of CD31. After culturing, the method includes transplanting the cultured cells into the individual for treatment purposes. The document also describes evidence that such BAT progenitor cells can differentiate into brown adipocytes characterized by high UCP1 expression.
The document describes differentiation and evaluation of the selected BAT progenitors, including the use of differentiation media and agents associated with induction of UCP1. Evidence in the provided content includes increased expression of mitochondrial and thermogenic markers and uncoupled respiration/proton leak, and includes a screening strategy using isolated CD34+ cells and a UCP1 promoter/enhancer reporter construct. The document also reports that UCP1 mRNA can be increased in muscle of rosiglitazone-treated obese type 2 diabetes patients.
Claims Coverage
The partial content includes two independent claims. Across the independent claims, there are at least two core inventive features: antibody-based identification of BAT progenitors from human skeletal muscle vascular cells using CD34/CD31 marker status, and a treatment method that cultures CD34+ / substantially CD31− BAT progenitor cells obtained from human skeletal muscle and transplants the cultured cells into the individual.
Antibody-based identification of CD34+ substantially CD31− BAT progenitors
providing a population of vascular cells isolated from human skeletal muscle; contacting the population of vascular cells with a first and a second antibody specific to CD34 and CD31 marker, respectively; and determining, in an antibody based assay, a BAT progenitor cell from the population of vascular cells that expresses the CD34 marker and is substantially free of the CD31 marker.
Culturing CD34+ substantially CD31− BAT progenitors and transplanting for treatment
culturing brown adipose tissue (BAT) progenitor cells obtained from human skeletal muscle of the individual or a donor in a medium, wherein said BAT progenitor cells are vascular cells that express CD34 marker and are substantially free of CD31 marker; and transplanting the cultured cells into the individual.
Overall, the independent claims center on defining BAT progenitor cells in human skeletal muscle vascular cells by antibody-based selection for CD34 expression with substantial absence of CD31, and using those CD34+ / substantially CD31− progenitors to treat metabolic disease by culturing them and transplanting the cultured cells into the individual.
Stated Advantages
Enables identification of BAT progenitor cells from vascular cells isolated from human skeletal muscle using an antibody based assay based on CD34 and substantially free of CD31.
Supports differentiation of BAT progenitor cells into brown adipocytes with high UCP1 expression, as described in the provided content.
Provides a method for treating metabolic disease in an individual by culturing CD34+ substantially CD31− BAT progenitor cells and transplanting the cultured cells.
Documented Applications
Identifying BAT progenitor cells from vascular cells isolated from human skeletal muscle using antibody-based assays with CD34 and substantially free of CD31.
Treating a metabolic disease or condition in an individual by culturing BAT progenitor cells from human skeletal muscle (from the individual or a donor) and transplanting the cultured cells into the individual.
Using isolated CD34+ cells and a UCP1 promoter/enhancer reporter construct for screening assays, as described in the provided content.
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